A comparison of nested polymerase chain reaction and immunofluorescence for the diagnosis of respiratory infections in children with bronchiolitis, and the implications for a cohorting strategy
A comparison of nested polymerase chain reaction and immunofluorescence for the diagnosis of respiratory infections in children with bronchiolitis, and the implications for a cohorting strategy
复制标题
DOI：
10.1053/jhin.2001.1044
复制
发表时间：
2001-10-01
影响因子：
6.9
通讯作者：
Coyle, PV
中科院分区：
文献类型：
--
作者：
Ong, GM;Wyatt, DE;Coyle, PV
对感染呼吸道合胞病毒（RSV）和/或流感病毒的毛细支气管炎患者进行队列检查，对于预防医院内这些病毒的交叉感染至关重要。采用巢式聚合酶链反应（nPCR）与免疫荧光法（IF）检测疑似毛细支气管炎患儿RSV A、B亚型。利用分子技术还研究了甲型流感（H3N2）、衣原体和小核糖核酸病毒/鼻病毒的联合感染。对2000年1月收治的毛细支气管炎患儿收集的50份鼻咽分泌物，包括RSV阳性（N=27）和RSV阴性（N= 23）标本，采用新聚合酶链反应（nPCR）检测两种RSV亚型，即甲型流感（H3N2）、衣原体和小核糖核酸病毒/鼻病毒。巢式PCR检测到28份RSV阳性标本（RSV A = 20份，RSV B = 8份），比IF法检测到的多2份，其中3份检测到甲型流感（H3N2）， 1份检测到沙眼衣原体，11份检测到小核糖核酸病毒，其中9份经巢式PCR确认为鼻病毒。nPCR检测双重感染5例。巢式PCR可用于检测呼吸道合胞病毒（RSV）和甲型流感（H3N2）感染，以及其他未常规调查的呼吸道病原体。由于不敏感技术的结果不准确，临床意义和潜在毒性病毒交叉感染的风险突出表明，需要将nPCR等分子测定作为常规调查方法，作为实验室服务的一部分提供。临床毛细支气管炎患者的队列应继续，同时等待实验室确认。(C) 2001医院感染学会。
Cohorting bronchiolitis patients infected with respiratory syncytial virus (RSV) and/or influenza viruses is paramount in preventing cross-infection of these viruses in hospital. Nested polymerase chain reaction (nPCR) was compared with immunofluorescence (IF) for the detection of RSV subtypes A and B in children with suspected bronchiolitis. Co-infection with influenza A(H3N2), Chlamydia spp. and picornavirus/rhinovirus was also investigated using molecular techniques.A total of 50 nasopharyngeal secretions collected from babies admitted with bronchiolitis in the month of January 2000, comprising IF RSV positive (N=27) and RSV negative (N= 23) specimens, were tested for both RSV subtypes, influenza A(H3N2), Chlamydia spp. and picornavirus/rhinovirus by nPCR.Nested PCR detected 28 specimens positive for RSV (RSV A = 20, RSV B = 8), which was two more than detected by IF Influenza A(H3N2) was detected in three specimens, Chlamydia trachomatis in one, and picornavirus in 11, of which nine were confirmed to be rhinovirus by nPCR. Dual infection was detected in five cases using nPCR.Nested PCR proved useful in detecting RSV and influenza A(H3N2) infections missed by IF, and also other respiratory tract pathogens not routinely investigated. The clinical implications and risk of cross-infection with potentially virulent viruses due to inaccurate results from insensitive techniques, highlights the need for molecular assays such as nPCR to be employed as a routine method of investigation, provided as part of the laboratory service. Cohorting of patients with clinical bronchiolitis should continue, whilst awaiting laboratory confirmation. (C) 2001 The Hospital Infection Society.